KR20100096788A - Pharmaceutical composition for preventing and treating diabetes, cancer or neurodegenerative diseases comprising boehmeria nivea extract as effective component - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing Boehmeria extract for preventing and treating diabetes, cancer, or neurodegenerative diseases is provided to suppress alpha-glucosidase, beta-glucosidase, beta-galactosidase, acethylcholine esterase(AChE), and butyrylcholine esterase(BChE). CONSTITUTION: A pharmaceutical composition for preventing and treating diabetes, cancer, or neurodegenerative diseases contains Boehmeria extract as an active ingredient. The Boehmeria includes Boehmeria nivea, Boehmeria nipononivea, Boehmeria spicata, Boehmeria tricuspis, Boehmeria sieboldiana, Boehmeria pannnosa, Boehmeria longispica, and Boehmeria platnifolia. A pharmaceutical composition for diabetes, cancer, or neurodegenerative disease contains the Boehmeria root butanol soluble extract or n-hexane soluble extract as an active ingredient.

Description

Pharmaceutical composition for preventing and treating diabetes, cancer or neurodegenerative diseases comprising Boehmeria nivea extract as effective component}

The present invention relates to a pharmaceutical composition and health functional food for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases containing the plant extract of the genus Fructus as an active ingredient.

Nettle ( Boehmeria nivea ( B. nivea ); common name: Rami, Germa ) belonging to the nettle family ( Urticaceae ) is one of the oldest and toughest weaving crops mainly used in textile manufacture and is widely used in subtropical and tropical Asia. Distributed over the area. Ramifula has also traditionally been used as a diuretic, antipyretic, hepatoprotective, antioxidant and anti-inflammatory agent. His fresh roots are ground into thick substances and used as a poultice (Lin et al., 1998. J Ethnopharmacol. 60, 9-17).

Matsuura's interest in the search for plant extracts has been described by Matsuura as Behenic Acid, Palmitic Acid, Stearic Acid, Ursolic Acid, 19α-hydroxyursolic Acid, β-Sitosterol, β-Sitosteryl-β-D-Glucoside, and Dow It began in 1973 when Kosterol et al. (Matsuura et al., 1973. Franch. Et Sav. Yakugaku Zasshi. 93, 1682-1684). Thereafter, the prospect of its use as a folk medicine continued, and the plant, despite its worth, was not concentrated on plant extracts and pharmaceutical research. With this in mind for the first time, we studied the glycosidase, cholinesterase, elastase and tyrosinase inhibitory properties of various plant parts of Mossipula, which are potential agents in many areas of disease control and treatment. It is worth it.

Oxidative stress is associated with the pathogenesis of various chronic diseases such as diabetes, cardiovascular disease and cancer. Antioxidants protect against free radicals, and therefore they are essential in achieving and maintaining good health. Much attention has been paid to polyphenols with strong antioxidant activity because they are effective in preventing the disease. Thus, DPPH free radical scavenging activity and total phenolic content were performed for all fractions (Zheng and Wang, 2001. J. Agric. Food Chem. , 49, 5165-5170).

Glycosidase inhibitors play an important role in the treatment of type II diabetes, viral or bacterial infections, lysosomal accumulation disorders and cancer, in which glycosidase is digested, biosynthesis of glycoproteins and lysosomal catabolism of glucoconjugates Because it is related to. Current symptomatic medicaments for the treatment of these diseases have a number of adverse / side effects, and are currently searching for alternative or traditional drug sources throughout the world (Mehta et al., 1998. FEBS Lett. 430, 17-22). ).

Cholinesterase inhibitors are the only approved drugs for the treatment of disorders associated with mild to severe Alzheimer's disease (AD), progressive degeneration of memory and cognitive function. The choline action hypothesis assumes that memory impairment in patients with Alzheimer's disease is due to a lack of choline action in the brain. Acetylcholinesterase inhibitors can restore levels of acetylcholine by inhibiting AChE. Efficient sources of AChE inhibitors are naturally provided from abundant plants in nature (Lopez et al., 2002. Life Sci. 72, 2521-2529). Therefore, interest is focused on finding novel cholinesterase inhibitors from natural sources, since only a few of these drugs are available for clinical use.

Korean Patent Laid-Open No. 2008-82955 discloses a pain-relieving composition comprising an underground extract of ramie grass as an active ingredient, and containing a pharmaceutically acceptable carrier, diluent or excipient.

The present invention has been made by the above-mentioned demands, and the present inventors have found that α-glucosidase, β-glucosidase, α-galactosidase, β-galactosidase, By examining the inhibitory activity of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes, we intend to use the ramie pool extract as a prophylactic and therapeutic agent for diseases related to these enzymes.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases containing the plant extracts of the genus Fructus as an active ingredient.

The present invention also provides a health functional food for preventing and improving diabetes, cancer or neurodegenerative diseases containing the extract as an active ingredient.

According to the present invention, the ramie pool extract of the present invention is α-glucosidase, β-glucosidase, β-galactosidase, acetylcholinesterase (AChE) and butyrylcholinesterase (depending on the ramie pool site and the solvent used). BChE). Thus, ramie grass extract can be used as a symptomatic or therapeutic agent for diabetes, cancer or neurodegenerative diseases related to the enzyme.

In order to achieve the object of the present invention, the present invention is Boehmeria nivea , Boehmeria nipononivea , Boehmeria spicata , Boehmeria spicata , Turtle tail ( Boehmeria tricuspis ), Long- leafed grass ( Boehmeria sieboldiana ), Bodhi pannnosa ), Boehmeria longispica and Boehmeria platnifolia , a pharmaceutical composition for the prevention and treatment of diabetes, cancer or neurodegenerative diseases, comprising as an active ingredient a plant extract of any genus to provide.

The present invention also provides a pharmaceutical composition for inhibiting α-glucosidase for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases containing butanol soluble extract of the plant root of the genus Root.

The present invention also provides a pharmaceutical composition for inhibiting β-glucosidase for the prevention and treatment of diabetes, cancer or neurodegenerative diseases, containing as an active ingredient ethyl acetate soluble extract of the plant roots of the genus Fructus.

The present invention also provides a pharmaceutical composition for inhibiting β-galactosidase for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases containing methylene chloride soluble extract of the plant leaves of the genus Fructus or ethyl acetate soluble extract of the stem as an active ingredient. to provide.

The present invention also provides a pharmaceutical composition for inhibiting acetylcholinesterase (AChE) for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases containing n-hexane soluble extract of the plant leaves of the genus Fructus.

The present invention also provides a pharmaceutical composition for inhibiting butyrylcholinesterase (BChE) for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases, containing methylene chloride soluble extract of the plant stem of the genus S. mori or n-hexane soluble extract of the root as an active ingredient. To provide a composition.

The present invention also provides a health functional food for preventing and improving diabetes, cancer or neurodegenerative diseases, containing the plant extract of the genus Morpho grass as an active ingredient.

Hereinafter, the present invention will be described in more detail.

The plant extract of the genus Fructus includes water and C1 to C4 lower alcohols such as methanol, ethanol, butanol and the like, and a mixed solvent thereof, preferably an extract available in methanol.

In addition, the plant non-polar solvent soluble extract of the genus Morsiful includes an extract soluble in a non-polar solvent such as hexane, chloroform, methylene chloride or ethyl acetate.

In addition, the plant polar solvent soluble extract of the genus Fructus may include one extracted with a polar solvent such as acetone, butanol, ethanol, methanol or water, preferably butanol.

The crude plant extract, non-polar solvent soluble extract and polar solvent soluble extract may be used as the leaves, stems, root extract of the plant genus.

Crude plant crude extract, nonpolar solvent soluble extract and polar solvent soluble extract of the present invention can be obtained as follows using, for example, ramie grass.

After collecting and drying the ramie grass of the present invention, and then grinding it with a mill, it is finely powdered, and the volume of water up to 20 times the weight of ramie grass sample and C1 to C4 lower alcohols such as methanol, ethanol, butanol, etc., or about 1: A mixed solvent having a mixing ratio of 0.1 to 1:10, preferably extraction with hydrothermal extraction, cold extraction, reflux cooling extraction or ultrasonic extraction for about 8 days at about 5 to 80 ℃, preferably about 24 ℃ with methanol Using the method, the supernatant may be recovered by vacuum filtration, and then the above procedure may be repeated 2 to 5 times, preferably twice, to collect the supernatant and concentrated under reduced pressure to obtain crude ramie extract.

In addition, the ramie pool non-polar solvent soluble extract of the present invention, after suspending the crude seed extract of distilled water in distilled water, about 1 to 100 times, preferably about 1 to 55 times the volume of hexane, ethyl acetate, chloroform A nonpolar solvent soluble layer may be extracted and separated by adding a nonpolar solvent 1 to 10 times, preferably 2 to 5 times. It is also possible to further carry out conventional fractionation processes (Harborne J. B. Phytochemical methods: A guide to modern techniques of plant analysis, 3rd Ed., Pp 6-7, 1998).

More preferably, the crude ramie extract obtained by the above process, preferably an organic solvent such as n-butanol, hexane, ethyl acetate, etc. is added in order from the lower polar solvent to the higher polar solvent, Hexane, ethyl acetate, n-butanol, water in this order sequentially solvent fractions, concentrated under reduced pressure to obtain a fraction of ramie hexane, ethyl acetate, n-butanol.

The present invention provides a pharmaceutical composition for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases, comprising as a active ingredient a plant extract of the genus Fructus, a nonpolar solvent-soluble extract, and a polar solvent-soluble extract containing the crude extract of the plant extract as an active ingredient. to provide.

The neurodegenerative diseases may include Alzheimer's disease, Parkinson's disease, Lou Gehrig's disease, Huntington's disease, and the like.

In addition, the plant of the genus of ramie, including ramie grass is a drug that has been used as a herbal medicine for a long time, the extracts of the present invention extracted therefrom also have no problems such as toxicity and side effects.

Moshiful methanol extract and each solvent fraction obtained by the above method inhibit α-glucosidase, β-glucosidase, β-galactosidase, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) Could confirm.

The pharmaceutical composition for preventing and treating diabetes mellitus, cancer or neurodegenerative disease of the present invention may include 0.02 to 50% by weight of the extract based on the total weight of the composition.

Pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

Pharmaceutical compositions comprising extracts according to the invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate and sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The extract of the present invention can be administered to mammals such as rats, mice, livestock, humans and the like in various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

In addition, the present invention provides a health functional food comprising the plant extract of the genus Fructus and food acceptable food supplements exhibiting a prevention and improvement effect of diabetes, cancer or neurodegenerative diseases. Examples of the food to which the extract can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to food or beverages for the purpose of preventing and improving diabetes, cancer or neurodegenerative diseases. At this time, the amount of the extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5 g, preferably 0.3 to 1g based on 100 ml. have.

The health functional beverage composition of the present invention has no particular limitation on the other ingredients other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract, for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. . The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the extract of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the extracts of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These ingredients may be used independently or in combination, and the proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example

1. Plant materials

The leaves, stems and roots of the ramie grass were obtained from the Korean biotech company "Korean Collection of Herbal Extracts." A collection of corroborating samples is available from these companies (Korea Collection of Herbal Extracts, 2000).

2. Extract

The dried leaves, stems and roots (2 Kg, dry mass for each plant site) were sliced into small pieces and kept at room temperature for 8 days in 100% methanol for intensive decoction. The extract was then concentrated at 20-30 ° C. using a rotary vacuum evaporator to yield a dried crude extract corresponding to leaves (70 g), stems (61 g) and roots (52 g).

3. Fractionation

Crude methanol extracts (70 g, 61 g and 52 g, respectively) of the leaves, stems and roots are suspended in distilled water (1 L), respectively, and partitioned into n-hexane, methylene chloride, ethyl acetate and n-butanol n -Hexane (22 g, 8 g and 11 g respectively), methylene chloride (3 g, 11 g and 4 g respectively), ethyl acetate (8 g, 14 g and 7 g respectively), n-butanol (each , 17 g, 10 g and 18 g) and water (16 g, 8 g and 22 g, respectively) fractions were obtained, respectively. Enzyme inhibitory activity assays were performed at 1 mg / ml concentration for the crude extract and all fractions thereof.

4. Reagents

α-glucosidase (from Saccharomyces cerevisiae type I), β-glucosidase (from almonds), α-galactosidase (from coffee beans), β-galactosida An aze (from Escherichia coli ), 4-nitrophenyl α-D-glucopyranoside, 4-nitrophenyl β-D-glucopyranoside, 4-nitrophenyl α-D-galactopyranoside, 2-nitrophenyl β-D-galactopyranoside, AChE (type VI-S from electric eel), BChE (from horse serum), acetylthiocholine iodide (ATCI), butyrylthiocholine iodide (BTCI), 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB), 1,1-diphenyl-2-picrylhydrazyl (DPPH), gallic acid and pauline-siokalto Folin-Ciocalteu reagent was purchased from Sigma-Aldrich (St. Louis, MO, USA). Other commercially available reagents and solvents were obtained and used.

5. Total Phenolic Content

The TPC of the extracts was determined using Pauline-Siocalto analysis with minor modifications to Zhang et al. (Zhang et al., 2006. J. Appl. Phycol. 18, 445-450). In this assay, 10 μl of each sample solution (concentration: 1 mg / ml) and 100 μl of Pauline-Siocalto reagent are mixed well, allowed to stand for 5 minutes, and then 80 μl of 7.5% sodium carbonate solution is added and Mixed. The covered plate was kept in the dark at room temperature for 30 minutes, and then the absorbance was measured at 750 nm using a spectrophotometer microplate reader. TPC was expressed as mg of gallic acid equivalent (GAE) per gram of dry extract.

6. DPPH Free radical scavenging activity

The free radical scavenging activity of the extracts was evaluated by a slight modification to the previously described method of Blois (Blois, M. S., 1958. Nature 181, 1199-1200). In this method, 50 μl of a methanol solution of DPPH (0.5 mM) was mixed with 10 μl of the test sample (concentration: 1 mg / ml) and 50 μl of 0.1 M Tris HCl buffer (pH 7.0), and the absorbance difference was reduced to 517 after 30 minutes. Measured at nm. The read value was compared with the control which contains 10 microliters of methanol instead of a test sample. All experiments were performed in 3 multiples.

7. Glucosidase Inhibition Assay

Methods previously reported by Shibano et al. (Shibano et al., 1997. Chem Pharm for enzyme inhibitory activity against α-glucosidase, β-glucosidase, α-galactosidase and β-galactosidase) Bull, 45, 700-5) was evaluated according to the method with some modification.

1) α-glucosidase assay

The reaction mixture was prepared with 50 μl 0.1 M phosphate buffer (pH 7.0), 25 μl 0.5 mM 4-nitrophenyl α-D-glucopyranoside (dissolved in 0.1 M phosphate buffer, pH 7.0), 10 μl test sample ( Concentration: 1 mg / ml) and 25 μl of α-glucosidase solution (immediately before analysis, 1 mg / ml stock solution in 0.01 M phosphate buffer at pH 7.0 using 0.1 Unit / diluted in ml). The reaction mixture was then incubated at 37 ° C. for 30 minutes. Subsequently, the reaction was finished by adding 100 µl of 0.2 M sodium carbonate solution. Enzymatic hydrolysis of the substrate was monitored by measuring the amount of p-nitrophenol released in the reaction mixture at 410 nm using a microplate reader. All experiments were performed in 3 multiples.

2) β-glucosidase assay

The reaction mixture was prepared with 50 μl 0.1 M phosphate buffer (pH 7.0), 25 μl 0.5 mM 4-nitrophenyl β-D-glucopyranoside (dissolved in 0.1 M phosphate buffer, pH 7.0), 10 μl test sample ( Concentration: 1 mg / ml) and 25 μl of β-glucosidase solution (immediately, 1 mg / ml stock solution in 0.01 M phosphate buffer at pH 7.0 with 2.5 Units / ml with the same buffer at pH 7.0). Dilution with). The reaction mixture was then incubated at 37 ° C. for 30 minutes. Subsequently, the reaction was finished by adding 100 µl of 0.2 M sodium carbonate solution. Enzymatic hydrolysis of the substrate was monitored by measuring the amount of p-nitrophenol released in the reaction mixture at 410 nm using a microplate reader. All experiments were performed in 3 multiples.

3) α-galactosidase assay

The reaction mixture is 50 μl 0.5 M phosphate buffer (pH 6.5), 25 μl 0.5 mM 4-nitrophenyl α-D-galactopyranoside (dissolved in 0.5 M phosphate buffer, pH 6.5), 10 μl test sample. (Concentration: 1 mg / ml) and 25 μl of α-galactosidase solution (just before analysis, 1 mg / ml stock solution in 50 mM phosphate buffer at pH 6.5, 0.1 Units / in the same buffer at pH 6.5). diluted in ml). The reaction mixture was then incubated at 37 ° C. for 30 minutes. Subsequently, the reaction was finished by adding 100 µl of 0.2 M sodium carbonate solution. Enzymatic hydrolysis of the substrate was monitored by measuring the amount of p-nitrophenol released in the reaction mixture at 410 nm using a microplate reader. All experiments were performed in 3 multiples.

4) β-D-galactosidase assay

The reaction mixture was prepared with 50 μl 0.1 M phosphate buffer (pH 7.3), 25 μl 0.5 mM 2-nitrophenyl β-D-galactopyranoside (dissolved in 0.1 M phosphate buffer, pH 7.3), 10 μl test sample. (Concentration: 1 mg / ml) and 25 μl of β-galactosidase solution (immediately before analysis, 1 mg / ml stock solution in 0.01 M phosphate buffer at pH 7.3, 0.44 Units / in the same buffer at pH 7.3). diluted in ml). The reaction mixture was then incubated at 37 ° C. for 30 minutes. Subsequently, the reaction was finished by adding 100 µl of 0.2 M sodium carbonate solution. Enzymatic hydrolysis of the substrate was monitored by measuring the amount of o-nitrophenol released in the reaction mixture at 410 nm using a microplate reader. All experiments were performed in 3 multiples.

8. AChE and BChE Inhibition Assay

Enzyme inhibitory activity against AChE and BChE was evaluated according to a method that had been slightly modified by the method previously reported by Ellman et al. (Ellman et al., 1961. Biochem Pharmacol. 7, 88-95). In this method, 20 μl of AChE solution (0.03 U / ml), 10 μl of test sample and 180 μl of buffer (pH 8) were mixed and incubated at 4 ° C. for 30 minutes. Subsequently, 20 μl of DTNB (0.3 mM) and 20 μl of ATCI (1.8 mM) were added to the reaction mixture, and after incubation at 37 ° C. for 20 minutes, the absorbance was measured at 412 nm and the percentage inhibition was calculated. Evaluation of BChE inhibition was performed as described above except the final enzyme concentration was 0.1 U / ml and the ATCI was replaced with BTCI (0.5 mM). Each sample was analyzed three times.

9. Statistical Analysis

All analyzes were performed three or more times using three multiples of samples. Inhibition rates for all assays were calculated as percentage of control (buffer with MeOH). All results were mean ± S.D. Marked as.

Example 1: Total Phenolic Content (TPC) and DPPH  ^. Free radical scavenging activity

As shown in Table 1, the ramie roots showed the best TPC (84 mg GAE / g) compared to the TPC of leaves and stems (29 and 48 mg GAE / g, respectively). At the root, after fractionation, significantly increased TPC was observed in the n-butanol and ethyl acetate fractions (190 and 180 mg GAE / g, respectively), whereas in all other fractions of leaves, stems and roots, TPC It was in the range of 1 to 80 mg GAE per gram of extract (Table 1).

Table 1. TPC of various plant parts of ramie grass and their respective fractions

Phenolic compounds are known to have antioxidant activity. The activity is believed to be mainly due to their redox properties, which play an important role in the absorption and neutralization of free radicals, the quenching of singlet and triplet oxygen or the decomposition of peroxides (Tepe et al., 2006. Food Chem. 95, 200-204).

Thus, the antioxidant activity of the different plant parts of the ramie grass is DPPH ^. It was measured using free radical scavenging analysis. Crude extract of root showed the best activity (66%) compared to other plant parts (Table 2). After fractionation, the n-butanol and ethyl acetate fractions showed significant activity (76% and 72%, respectively), which accounts for the high amount of correlation between total phenolic and antioxidant activity in ramie pool.

Table 2. Inhibition rates of various plant parts of ramie pools and their respective fractions in different assay systems

There is increasing evidence that free radicals and reactive oxygen species can induce oxidative damage in biomolecules such as lipids, proteins and nucleic acids, leading to human diseases such as atherosclerosis, cancer, diabetes and neurodegenerative planetary disorders. Herbal drugs containing such free radical scavengers are known for the treatment of such diseases (Beauchamp, C., Fridovich, I., 1971. Anal. Biochem. 44, 276-287).

In addition, various in vitro assays were performed to explore the potential of the Lamiaceae extract for treating the aforementioned diseases.

Example 2: Glycosidase Inhibition Assay

Glycosidase inhibitors are promising therapeutics for some degenerative diseases, including diabetes, viral attachment and cancer (Mehta et al., 1998. FEBS Lett. 430, 17-22). In the literature, a number of herbal extracts have been reported to be used as antidiabetic agents, but no such studies have been carried out on ramie pool extracts in the past. Thus, for the first time, mossipip crude extract (in 100% methanol) and its n-hexane, methylene chloride, ethyl acetate, n-butanol and aqueous fractions are α-glucosidase, β-glucosidase, α-galacto Cedarase and β-galactosidase inhibitory activity was investigated.

Compared to the leaf and stem extracts, fractions of the root extracts of the ramie grass showed significant α-glucosidase and β-glucosidase inhibitory activities in the range of 51 to 94% and 51 to 93%, respectively, as described in Table 2 above. It is. The highest inhibition was reported to be 94% and 93% α-glucosidase and β-glucosidase inhibition in the n-butanol and ethyl acetate fractions, respectively.

β-galactosidase inhibition ranged from 2 to 95% in leaf extracts and methylene chloride showed 95% inhibition. In stem extracts, significant inhibition was observed in the ethyl acetate fraction (77%), but no inhibition was found in root extracts for β-galactosidase (Table 2). In all the extracts of the ramie grass, little α-galactosidase inhibitory activity was shown.

In summary, mociful is an effective biosource for α-glucosidase, β-glucosidase and β-galactosidase inhibitory activity.

Example 3: AChE and BChE Inhibition Assay

Since AD, one of the most common causes of death worldwide, is a threat to public health, there has been a growing interest in new therapeutic strategies based on medicinal plants. The efficacy of the ramie grass extract was assessed using the AChE and BChE assays. The results obtained in this study suggested that the stem and root extracts of the ramie grass were potent BChE inhibitors, where the methylene chloride fraction of the stem and the n-hexane fraction of the root showed 74% and 66% inhibition, respectively. Moderate AChE inhibition was observed in the n-hexane fraction (50%). AChE and BChE inhibition in all fractions of various parts of the ramie pool ranged from 0 to 50% and 27 to 74%, respectively (Table 2). This indicates that the ramie grass extract may prove to be a potential candidate for the treatment of AD and other neurodegenerative diseases such as Parkinson's disease, senile dementia, dyskinesia and myasthenia gravis.

Hereinafter, an example of the preparation of a pharmaceutical composition including the extract of the present invention will be described, but the present invention is not intended to limit the present invention, but is intended to be described in detail.

Formulation Example 1 Preparation of Powder

Ramie Grass Extract Powder 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2: Preparation of Tablet

Ramie grass extract powder 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3: Preparation of Capsule

Ramie grass extract powder 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

Ramie grass extract powder 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

_{Na 2 HPO 4 · 12H 2 O} 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Ramie Grass Extract Powder 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

According to the conventional method for preparing a liquid, each component is added and dissolved in purified water, lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Drinks

Ramie Grass Extract Powder 100mg

15 g of vitamin C

100 g of vitamin E (powder)

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B ₁ 0.25 g

Vitamin B ₂ 0.3 g

Water quantification

After mixing the above components according to the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilized and then refrigerated and stored Used to prepare the healthy beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (7)

Ramie (Boehmeria nivea), Island ramie (Boehmeria nipononivea), jomkkae ipnamu (Boehmeria spicata), Turtle Tail (Boehmeria tricuspis), ginip ramie (Boehmeria sieboldiana), king of ramie (Boehmeria pannnosa), why ramie (Boehmeria longispica) and more ramie ( Boehmeria platnifolia ) A pharmaceutical composition for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases, containing as an active ingredient any one of the plant extracts of the genus Fructus selected from the group consisting of. The pharmaceutical composition for inhibiting α-glucosidase for preventing and treating diabetes mellitus, cancer or neurodegenerative disease according to claim 1, comprising butanol soluble extract of the root of the plant of genus Fructus. The pharmaceutical composition for inhibiting β-glucosidase for preventing and treating diabetes mellitus, cancer or neurodegenerative disease according to claim 1, comprising the ethyl acetate soluble extract of the plant roots of the genus Fructus as an active ingredient. The pharmaceutical composition for inhibiting β-galactosidase for preventing and treating diabetes mellitus, cancer, or neurodegenerative disease according to claim 1, comprising the methylene chloride soluble extract of the plant leaves of the genus Fructus or the ethyl acetate soluble extract of the stem as an active ingredient. Composition. The pharmaceutical composition for inhibiting acetylcholinesterase (AChE) for preventing and treating diabetes mellitus, cancer or neurodegenerative disease according to claim 1, comprising n-hexane soluble extract of the plant leaves of the genus Fructus. According to claim 1, Butyryl cholinesterase (BChE) inhibition for the prevention and treatment of diabetes mellitus, cancer or neurodegenerative diseases containing methylene chloride soluble extract of the plant stem of the genus Fructus or n-hexane soluble extract of the root as an active ingredient Pharmaceutical composition for. A health functional food for preventing and improving diabetes, cancer or neurodegenerative diseases, comprising the plant extract of the genus Fructus of any one of claims 1 to 6 as an active ingredient.